Deep Dermatophytosis Caused by Zoophilic Strain of <Emphasis Type="Italic">Trichophyton interdigitale</Emphasis> with Successful Treatment of Itraconazole

We report a 66-year-old female patient with deep dermatophytosis caused by zoophilic strain of Trichophyton interdigitale, a rare granulomatous presentation of Trichophyton species infection in patients with underlying systemic diseases, and she was successfully cured by itraconazole. Since the identification of Trichophyton mentagrophytes complex had been misused for years, a brief discussion of molecular diagnosis and taxonomy of T. mentagrophytes complex is given. The pathogenesis and comparison with cases reported in the literature are also discussed.     

Cryopreservation of hairy root cultures of <Emphasis Type="Italic">Maesa lanceolata</Emphasis> and <Emphasis Type="Italic">Medicago truncatula</Emphasis>

To study the production of secondary metabolites of Maesa lanceolata and Medicago truncatula, hairy root cultures of both plant species were established. Because maintenance of large numbers of cultures is laborious and costly, we developed a cryopreservation protocol and stored different isolated lines over time. Using encapsulation-dehydration, high survival rates were observed for both Maesa and Medicago hairy roots. Root tips were isolated and encapsulated in calcium-alginate beads, containing 0.1 M sucrose. The encapsulated hairy roots were precultured for 3 days using basal medium containing high sucrose concentrations. Medicago root tip growth during the preculturing time lead to unwanted outgrowth which could be tempered by addition of plant growth inhibitors. After preculturing, the beads were dehydrated in the air flow of a laminar flow until 35–40% of the initial bead weight was reached. Dehydrated beads were plunged into liquid nitrogen and after different storage times thawed in a water bath at 40°C. The survival rates were 90% for Maesa and 53% for Medicago, which are sufficient to allow implementation in large storage experimental set-ups.     

Association of methylenetetrahydrofolate reductase gene 677C > T polymorphism and Down syndrome

The association between Down syndrome (DS) and maternal polymorphisms in genes encoding folic acid metabolizing enzymes remains a controversial issue. A meta-analysis was performed to evaluate the association of maternal MTHFR 677C > T polymorphism and the risk of having a child with DS. Case–control studies were screened from major literature databases. Twenty articles from 13 countries worldwide, with a total of 2,101 DS and 2,702 control mothers, attended the inclusion criteria. We found a 50 % increase for the association of maternal homozygous TT genotype and DS in both fixed (OR = 1.51; 95 % CI 1.22–1.87) and random effects models (OR 1.54; 95 % 1.15–2.05). Similarly, a significant pooled OR was found for the heterozygote CT, with an OR 1.26; 95 % CI 1.10–1.43 (fixed effects model) and OR 1.28; 95 % 1.08–1.51 (random effects model). As ultra-violet B solar radiation highly depends on latitude, and can promote, in less pigmented skin, intravascular folate photolysis, we stratified the analysis by latitude region, defining as Tropical (between 23.5^° S and 23.5^° N), Sub-Tropical (between 23.5^° and 40^° N and S), and Northern (≥40^o N). Significant association was only found for Sub-Tropical area, both using fixed and random effect models. In conclusion, MTHFR 677C > T polymorphism is a moderate risk factor for DS for some populations, and populations located in Sub-Tropical region seem to be at greater risk. Latitude, ethnicity, skin pigmentation, and red blood cell folate are important variables to be considered in future studies.     

Anthraquinones from in vitro root culture of <Emphasis Type="Italic">Morinda royoc</Emphasis> L.

Morinda royoc L. (Rubiaceae) root cultures were established for the production of anthraquinones (AQs). Three independent experiments were performed to evaluate the effects of different levels of indole-3-acetic acid (0–22.8 μM), culture duration (15–75 days) and subculture number (0–4). The following indicators were recorded: root fresh weight per Erlenmeyer and intracellular and extracellular AQ production. The experiments performed in this study allowed an increase of intracellular AQ content up to a maximum of 4.5 mg g⁻¹ of fresh mass, after 30 days of culture in a medium 5.7 μM of IAA. In addition, isolation and identification of seven AQs from M. royoc L. roots is described, one of them being reported for the first time for this species. The structures of isolated compounds were determined from ¹H-NMR data. To the best of our knowledge, this is the first report on AQ production from root culture of this plant.     

Transgenic expression of the von Willebrand A domain of the BONZAI 1/COPINE 1 protein triggers a lesion-mimic phenotype in<Emphasis Type="Italic"> Arabidopsis</Emphasis>

The copines are a newly identified, widely distributed class of Ca²⁺-dependent, phospholipid-binding proteins that may be involved in cellular signaling. The copines have a characteristic domain structure: two C2 domains in the N-terminal region and a von Willebrand A (VWA) domain in the C-terminal region. Studies suggest that copines interact with target protein(s) via their VWA domain and recruit the proteins to a membrane location through the activity of the C2 domains. Arabidopsis thaliana (L.) Heynh. plants with loss-of-function mutations in the BONZAI 1/COPINE 1 (BON1/CPN1) gene display aberrant regulation of defense responses, including development of a lesion-mimic phenotype, an accelerated hypersensitive response, and increased resistance to a bacterial and an oomycetous pathogen. The phenotype of mutants in BON1/CPN1 is both humidity- and temperature-sensitive. In this study, we generated transgenic plants expressing either the VWA or the C2 portions of BON1/CPN1 in the wild-type Columbia-0 (Col-0) genetic background. Transgenic plants expressing the BON1/CPN1 C2 domain portion appeared like wild-type plants. However, transgenic plants expressing the BON1/CPN1 VWA domain exhibited a lesion-mimic phenotype that partially phenocopied bon1/cpn1 mutant plants. Our data suggest that BON1/CPN1 VWA domain fragments may interfere with the function of the full-length endogenous BON1/CPN1 protein, possibly by competing with the full-length BON1/CPN1 protein for association with target proteins normally bound to the full-length BON1/CPN1 protein.     

<Emphasis Type="Bold">The Dangers of Direct-to-Consumer Genetic Testing for Alzheimer’s Disease</Emphasis>

The overarching issue with this case study is poor regulation and quality control over direct-to-consumer genetic testing, delivered in the absence of any medical oversight.     

Mapping of the chromosome 17 BMD QTL in the F<Subscript>2</Subscript> male mice of MRL/MpJ × SJL/J

Developing treatment strategies for osteoporosis would be facilitated by identifying genes regulating bone mineral density (BMD). One way to do so is through quantitative trait locus (QTL) mapping. However, there are sex differences in terms of the presence/absence and locations of BMD QTLs. In a previous study, our group identified a BMD QTL on chromosome 17 in the F₂ female mice of the MRL/MpJ × SJL/J cross. Here, we determined whether it was also present in the male mice of the same cross. Furthermore, we also intended to reduce the QTL region by increasing marker density. Interval mapping showed that the same QTL based on chromosomal positions was present in the male mice, with logarithmic odds (LOD) scores of 4.0 for femur BMD and 5.2 for total body BMD. Although there was a body weight QTL at the same location, the BMD QTL was not affected by the adjustment for body weight. Mapping with increased marker density indicated a most likely region of 35–55 Mb for this QTL. There were also co-localized QTLs for femur length, femur periosteal circumference (PC) and total body bone area, suggesting possibility of pleiotropy. Runx2 and VEGFA are strong candidate genes located within this QTL region.     

Conditions for Global Dynamic Stability of a Class of Resource-Bounded Model Ecosystems

This paper studies a class of dynamical systems that model multi-species ecosystems. These systems are ‘resource bounded’ in the sense that species compete to utilize an underlying limiting resource or substrate. This boundedness means that the relevant state space can be reduced to a simplex, with coordinates representing the proportions of substrate utilized by the various species. If the vector field is inward pointing on the boundary of the simplex, the state space is forward invariant under the system flow, a requirement that can be interpreted as the presence of non-zero exogenous recruitment. We consider conditions under which these model systems have a unique interior equilibrium that is globally asymptotically stable. The systems we consider generalize classical multi-species Lotka–Volterra systems, the behaviour of which is characterized by properties of the community (or interaction) matrix. However, the more general systems considered here are not characterized by a single matrix, but rather a family of matrices. We develop a set of ‘explicit conditions’ on the basis of a notion of ‘uniform diagonal dominance’ for such a family of matrices, that allows us to extract a set of sufficient conditions for global asymptotic stability based on properties of a single, derived matrix. Examples of these explicit conditions are discussed.     

Gastric cancer & prospects of cancer in Saudi Arabia peninsula

Gastric cancer is classified to be an aggressive disease with poor treatment outcome, as most cases remain undetected until later stages, wherein surgery and few chemotherapeutics become the only recommended treatment course. The process of cancer development is multistep involving many stages and types of precancerous lesions, and hence, routine monitoring becomes a necessity in those detected with these or exposed to risk factors. Studying the pattern of gastric cancer for any geographical region is also important to control mortality and focus on implementation of efficient management and treatment guidelines. The cause for gastric cancer can be genetic, racial as well as environmental, and hence the pattern of this malignancy differs across geographical regions and between the developing and the developed nations. In case of the Kindgom of Saudi Arabia, very few hospital-based reports have been published highlighting the pattern of gastric cancer, and the associated incidence and mortality rates. However, classified to be one of the most crucial cancer forms in Saudi Arabia, research pertaining to epidemiology, presentation and pathological features are limited. Studying gastric cancer occurrence from public health viewpoint is important also because eradication of causative agents like those that H. pylori has also shown been not reduce the risk of cancer development among individuals with atrophic metaplastic gastritis. In case of Saudi Arabia, many inherent risks for this malignancy exists like waterpipe smoking and shift in diet pattern from the traditional Mediterranean diet. Our review focusses on pattern of gastric cancer on a global scale in comparison to scenario in Saudi Arabia. The aim is to encompass all of the less stressed upon facts about this malignancy in the Kingdom, paving way for future work in this regards.     

A linkage map of maize × teosinte <Emphasis Type="Italic">Zea luxurians</Emphasis> and identification of QTLs controlling root aerenchyma formation

The objectives of this study were to construct a linkage map and identify quantitative trait loci (QTLs) controlling root aerenchyma formation in drained soil conditions using 195 F₂ individuals derived from a cross between maize inbred line B73 × teosinte Zea luxurians. A 107 SSR marker based map covering 1,331 cM across all ten chromosomes was developed. One significant difference between the parents utilized in the study was that under non-flooding conditions, B73 exhibits a minor capacity to develop root aerenchyma, whereas Z. luxurians exhibits a high tendency to form aerenchyma. Linkage analysis indicated segregation distortion regions on chromosomes 2, 4 and 8, and severe recombination suppression on the long arm of chromosome 4. Multiple interval mapping analysis suggests that five QTLs for aerenchyma formation in non-flooding conditions are located on chromosomes 2, 3, 5, 9 and 10, and these explained 36.3% of the total phenotyphic variance. The Z. luxurians alleles in all five QTLs increased the capacity to form aerenchyma and the locations of these QTLs did not overlap those previously identified in the teosinte Z. nicaraguensis. By transferring aerenchyma-forming QTLs from both Z. luxurians and Z. nicaraguensis, it may be possible to pyramid these genes and develop a maize line with exceptional aerenchyma formation and a high level of tolerance to flooding conditions.     

Environmental Fluctuations and Level of Density-Compensation Strongly Affects the Probability of Fixation and Fixation Times

The probability of, and time to, fixation of a mutation in a population has traditionally been studied by the classic Wright–Fisher model where population size is constant. Recent theoretical expansions have covered fluctuating populations in various ways but have not incorporated models of how the environment fluctuates in combination with different levels of density-compensation affecting fecundity. We tested the hypothesis that the probability of, and time to, fixation of neutral, advantageous and deleterious mutations is dependent on how the environment fluctuates over time, and on the level of density-compensation. We found that fixation probabilities and times were dependent on the pattern of autocorrelation of carrying capacity over time and interacted with density-compensation. The pattern found was most pronounced at small population sizes. The patterns differed greatly depending on whether the mutation was neutral, advantageous, or disadvantageous. The results indicate that the degree of mismatch between carrying capacity and population size is a key factor, rather than population size per se, and that effective population sizes can be very low also when the census population size is far above the carrying capacity. This study highlights the need for explicit population dynamic models and models for environmental fluctuations for the understanding of the dynamics of genes in populations.     

Expression profiles of <Emphasis Type="Italic">amh</Emphasis> and <Emphasis Type="Italic">foxl2</Emphasis> in <Emphasis Type="Italic">Schizothorax kozlovi</Emphasis>, and their response to temperature during the early developmental stage

To elucidate the role of amh and foxl2 in sex differentiation of the teleost fish Schizothorax kozlovi, the full-length cDNAs were cloned from the mature testis and ovary by rapid amplification of cDNA ends (RACE), and their relative mRNA expression levels were determined by quantitative real-time polymerase chain reaction among tissues and temperature groups. The complete amh and foxl2 cDNAs of S. kozlovi were 2060 bp and 1750 bp, which encoded 568 and 306 amino acids, respectively. The amh were expressed only in gonads, while foxl2 was expressed in the gills, brain and gonads, both exhibiting relatively high tissue specificity. The amh exhibited sex-specific expression pattern in the gonads. No sex differences in the foxl2 expression were observed in the brain and gonads, but significant sex differences were found in the gills. No significant differences were found in the foxl2 expression, from the larval to the juvenile stage, and also between different temperature groups. However, significant differences were found in the expression levels of amh from the larval (12–63 days posthatching (dph)) to the juvenile stage (190 dph), and also among the \(18{^{\circ }}\hbox {C}\) and \(10{^{\circ }}\hbox {C}\) groups at 31 dph. This result suggested that amh plays an important role in male sex differentiation of S. kozlovi during the early developmental stage, but no similar effect was observed in foxl2.     

Invasion ecology of quagga mussels (<Emphasis Type="Italic">Dreissena rostriformis bugensis</Emphasis>): a review of evolutionary and phylogenetic impacts

Two invasive freshwater mussels, Dreissena rostriformis bugensis (quagga mussel) and D. polymorpha (zebra mussel), reveal differences in patterns and timing of their invasions in Europe. They belong to different clades in Dreissena phylogenetics: D. rostriformis bugensis genetically is coupled with the brackish water, lacustrine D. r. distincta and the two are believed to represent a single species. As such, the guaqqa mussel has environmental requirements that differ from the congeneric D. polymorpha. D. rostriformis bugensis invasions were confined to reservoirs of the Dnieper, Don and Dniester Rivers of the Black Sea basin. We recorded D. r. bugensis outside the Black Sea basin for the first time between 1992 and 2001, along the Volga River reservoir cascade including the Northern Caspian Sea shallows. This represents a 40-year invasion time lag since an invasion corridor through the Volgo-Don Waterway was established in 1952 (a corridor used extensively by many invertebrate species from the Black Sea region). We attribute the postponed invasion of Europe by D. r. bugensis, including peculiarities in establishment and its absence in fossil records, to its phylogenetically close relationship with D. r. distincta and its recent evolutionary origin. The relatively rapid range expansion of D. r. bugensis in eastern Europe during the past several decades was facilitated by human-mediated ecosystem transformation, notably impoundment of large eastern European rivers, that have allowed this species to utilize newly transformed ecosystems.     

Construction of a Coix BAC library and isolation of the 22 kDa &alpha;-coixin gene cluster

Coix lacryma-jobi L. (Coix) is a close relative of maize and is considered a valuable genetic resource for crop improvement. Here we report the construction of the first Coix bacterial artificial chromosome (BAC) library using accession PI 324059. This BAC library contains about 230?400 clones with an average insert size of 113?kb, has low organellar DNA contamination, and provides 16.3-fold coverage of the genome. The library was stored in 12?× 96 pools that could be screened with a PCR protocol. Library screening was performed for the 22?kDa α-coixin gene family. A total of 57 positive pools were identified, and single clones were isolated from 19 of these pools. Based on DNA fingerprinting and Southern blot analysis, these 19 BAC clones form a single contig of about 340?kb in length, indicating that the 22 kDa α-coixin genes occur in a cluster. These results demonstrated the suitability of this BAC library for gene isolation and comparative genomics studies of the Coix genome.     

Comparison of Monte Carlo Simulations of Cytochrome b<Subscript>6</Subscript>f with Experiment Using Latin Hypercube Sampling

We have programmed a Monte Carlo simulation of the Q-cycle model of electron transport in cytochrome b₆f complex, an enzyme in the photosynthetic pathway that converts sunlight into biologically useful forms of chemical energy. Results were compared with published experiments of Kramer and Crofts (Biochim. Biophys. Acta 1183:72–84, 1993). Rates for the simulation were optimized by constructing large numbers of parameter sets using Latin hypercube sampling and selecting those that gave the minimum mean square deviation from experiment. Multiple copies of the simulation program were run in parallel on a Beowulf cluster. We found that Latin hypercube sampling works well as a method for approximately optimizing very noisy objective functions of 15 or 22 variables. Further, the simplified Q-cycle model can reproduce experimental results in the presence or absence of a quinone reductase (Q_i) site inhibitor without invoking ad hoc side-reactions.     

Presence of 46 kDa Gelatinase on the Outer Membrane of <Emphasis Type="Italic">Leptospira</Emphasis>

Leptospira infection involves the adhesion of the bacteria followed by invasion of the host crossing the extracellular matrix barrier. In an effort to understand the molecular mechanism of this process, the possibility of occurrence of matrix degrading enzymes from Leptospira was investigated. Zymographic analysis showed that the outer membrane of Leptospires contains a gelatinase of average molecular size of 46 kDa. The gelatinase exhibited maximum activity at neutral pH and was inhibited by metal chelators such as EGTA, EDTA, and Orthophenanthroline and was activated by calcium, magnesium, zinc, and copper, suggesting that it is a membrane-associated neutral matrix metalloproteinase. Analysis of the production of the enzyme by various serovars showed that the pathogenic serovars expressed significant amount of this enzyme while nonpathogenic forms either did not express or showed only very low activity, suggesting that this enzyme may be associated with pathogenesis of leptospirosis.     

Assessment of antifungal effects of a novel compound from <Emphasis Type="Italic">Burkholderia cepacia</Emphasis> against <Emphasis Type="Italic">Fusarium solani</Emphasis> by fluorescent staining

A novel compound CF66I produced by Burkholeria cepacia was investigated for its antifungal effects against Fusarium solani by three different fluorescent dyes. Dual staining with propidium iodide (PI) and fluorescein diacetate (FDA) demonstrated high doses of CF66I (120.0 μg ml⁻¹) killed the fungi by acting primarily on the cell membrane. However, at fungistatic concentration (20.0 μg ml⁻¹) of this compound, microscopic observations revealed swelling hyphae with abnormal chitin deposition, as determined by Calcofluor white (CFW) staining, which was indicative of the alterations in cell wall structure. In addition, inhibition of intracellular esterases activity was observed. These results led us to conclude that low doses of CF66I probably inhibited the fungal growth by interfering with the cell metabolic pathways.     

Optimization of the isolation and cultivation of <Emphasis Type="Italic">Cyprinus carpio</Emphasis> primary hepatocytes

The aquatic environment is affected by numerous chemical contaminants. There is an increasing need to identify these chemicals and to evaluate their potential toxicity towards aquatic life. In this research we optimized techniques for primary cell culture of Cyprinus carpio hepatocytes as one adjunct model for ecotoxicological evaluation of the potential hazards of xenobiotics in the aquatic environment. In this study, Cyprinus carpio hepatocytes were isolated by mechanical separation, two-step collagenase perfusion, and pancreatin digestion. The hepatocytes or parenchymal cells could be separated from cell debris and from non-parenchymal cells by low-speed centrifugation (Percoll gradient centrifugation). The harvested hepatocytes were suspended in DMEM, M199 (cultured in 5% CO₂), or L-15 (cultured without 5% CO₂) medium then cultured at 17, 27, or 37 °C. Cell yield was counted by use of a hemocytometer, and the viability of the cells was assessed by use of the Trypan blue exclusion test. Results from these studies showed that the best method of isolation was pancreatin digestion (the cell yield was 2.7 × 10⁸ per g (liver weight) and the viability was 98.4%) and the best medium was M199 (cultured in 5% CO₂) or L-15 (cultured without 5% CO₂). The optimum culture temperature was 27 °C. The primary hepatocytes culture of Cyprimus carpio grew well and satisfied requirements for most toxicological experiments in this condition.     

Taxonomic implication of embryo micromorphology in the genus <Emphasis Type="Italic">Vicia</Emphasis> L. (Fabaceae)

In this work, embryos of selected Vicia species were examined to reveal the micromorphological characters that could have taxonomic significance. Experimental results show that morphological characters such as cotyledons shape and colour, radicle shape, colour and position, plumule colour and stipular parts attachments are variable and could be used for taxa identification in the genus Vicia. Numerical analysis based on 38 micromorphological characters was used to build a phenogram that indicated the relationships among the studied taxa. In addition, an identification key using embryo characters was prepared for Vicia species. The variability of embryo characters were discussed in the taxonomic context. Although the embryo characters have only limited phylogenetic signal for Vicia species, they might be used for their morphological delimitation.     

Isolation and characterization of butanol-tolerant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>

Objectives

A new solvent-tolerant species, Staphylococcus aureus, was isolated and characterized during the screening of butanol-tolerant microorganisms.

Results

Three isolates of S. aureus were obtained as contaminants during improvement of butanol tolerance of E. coli K12. Their cell dry weights were 135 % that of K12 in the absence of butanol stress. S. aureus had a growth advantage over K12 when cultured with various concentrations of butanol. It can tolerate up to 3 % (v/v) butanol, while most solventogenic bacteria can tolerate only 2 % (v/v) butanol. The addition of 10–20 g glucose/l enhanced its butanol tolerance. The relative cell biomass of the S. aureus was 71–306 % that of E. coli under 5.5–10 % (v/v) ethanol stress, indicating ethanol resistance.

Conclusions

This is the first study to observe butanol-tolerant S. aureus. As this organism can be genetically manipulated, it could have a wide array of applications.